Journal: Frontiers in Neural Circuits

Article Title: Intense Activity of the Raphe Spinal Pathway Depresses Motor Activity via a Serotonin Dependent Mechanism

doi: 10.3389/fncir.2017.00111

Figure Lengend Snippet: Unspecificity of available 5HT 1A antibodies. (A) HEK293 cells transfected with HA-tagged h5-HT 1A and stained with anti-HA antibodies in combination with five different anti-5-HT 1A antibodies. Anti-5-HT 1A staining (red) obtained using the five antibodies is depicted in the left column, while anti-HA (green) is shown in the middle column. Only two antibodies were able to recognize HA-h5-HT 1A expressing HEK293 cells, S1A-170 and H-119 from Santa Cruz Biotechnology. The remainder of the antibodies revealed either no staining or unspecific staining. Staining of the nuclei with 4′,6-diamidino-2-phenylindole (DAPI) is included in blue in the merged images to the right. (B) Turtle spinal cord section triple-labeled with S1A-170 (red), 5-HT (green) and choline acetyltransferase (ChAT, blue, marker of motorneurons). S1A-170 stained the proximal region on a motoneuron neurite consistent with the previously reported AIS staining with this antibody. (C) Mouse hippocampal CA1 wild-type and knockout tissue sections were stained with S1A-170 (green). As shown, stainings consistent with the localization of the AIS was observed in pyramidal cells of wild-type animals. The staining was, however, not eliminated in 5-HT 1A knockout animals demonstrating that this staining is not specific for 5-HT 1A . The right column represents overlay images of S1A-170 immunoreactivity and DAPI signal (blue).

Article Snippet: AB144P, Merck Life Science A/S), rabbit anti-5-HT 1A (1:100, H-119, Santa Cruz Biotechnology), rabbit anti-5-HT 1A (1:50, ASR-021, Alomone Labs), rabbit anti-5-HT 1A (1:100, ADI905-741-100, Enzo Life Sciences), rabbit 5-HT 1A (1:100, NB100-92418, Novus Biologicals), and mouse anti-HA epitope (1:500 dilution, clone 16B12, Biolegend).

Techniques: Transfection, Staining, Expressing, Labeling, Marker, Knock-Out